Diagnostic immunology最新文献

As part of a study to define the factors affecting the distribution of lymphocyte subpopulations in healthy blood donors, we have measured lymphocyte surface markers in various racial groups. Markers tested were T3 (all T cells), T4A (T helper cells), T8 (T suppressor cells), and Leu 11 (natural killer cells). Racial groups included three Asian groups (Chinese, Japanese, Other Orientals) and three non-Asian groups (Caucasians, Hispanics, American Blacks). The mean percentage of T3 + cells and T4A + cells were significantly lower in Asians compared with non-Asians, while T8 levels did not differ. These changes resulted in a significantly lower mean T4A:T8 ratio in Asians compared with non-Asians. The mean percentage of Leu 11 + cells was higher in Asians compared with non-Asians. Within the Asian group, Chinese had a higher mean Leu 11 + value than the other two Asian groups combined. Further, pairwise comparisons showed that Chinese had a significantly higher mean Leu 11 + value compared with each of the other five racial groups. This increased mean Leu 11 + level in the Chinese group reflected a distinct cluster of high values for about half the subjects. These findings show that racial background should be a major consideration when defining the normal range for lymphocyte subpopulations detected by surface markers.

Using a battery of monoclonal antibodies on snap-frozen sections, we delineated the immunoarchitecture of two splenic small cleaved cell lymphomas (SCL). Both cases had light- and heavy-chain restricted immunoglobulin (lg) expression signifying replacement of splenic white pulp by a single B-cell clone. Both the monotypia and aberrant topography of lg expression in SCL contrasted with the usual polyclonal, zonal lg expression in reactive splenic B-cell zones. Pan B antigens (B1, B4, and L14) were constant in expression as expected for B-cell neoplasms, while B-cell maturation antigens (B2, IgD, and CALLA) were variably expressed, suggesting that different SCL may derive from separate phases of B-cell ontogeny. Close association of SCL with dendritic reticulum cells suggests SCL may derive from splenic secondary follicles or home to these sites. The variable T-cell component detected by a T-cell panel (Leu 1-9) indicates the substantial range of T-cell reactivity in splenic SCL. We emphasize the immunologic aberrancy of splenic SCL when compared to normal splenic B-cell immunotopography. Further, we illustrate the utility of serial tissue section immunochemistry in revealing complex neoplastic cell phenotypes and in revealing the relationships of reactive cells to neoplastic cells.

Hybridoma AR1-28 [Chin and Miller, Cancer Res 45:1723-1729, 1985] was produced using splenocytes from BALB/c mice which had been immunized with human pancreatic adenocarcinoma cell line RWP-1 [Dexter et al, Cancer Res 42:2705-2714, 1982]. This antibody cross reacted with the RWP-2 cell line. AR1-28, an IgG1 antibody, stained the membranes of five (RWP-1, RWP-2, BxPC-3, HPAF-2, and T3M4) out of ten human pancreatic tumor cell lines by immunofluorescence techniques. Electron microscopy on RWP-2 cells, stained by indirect immunoperoxidase, confirmed a membrane location for the AR1-28 antigen. Twenty-three of the 27 clinical specimens (83%) of formalin-fixed, paraffin-embedded pancreatic cancers tested were positive. Varying intensities of staining were observed and were related to the degree of differentiation achieved by the tumor: poorly differentiated tumors showing the least staining while well-differentiated tumors showed the greatest intensity of staining in a predominantly apical location. Immunoblotting showed that AR1-28 reacts with a 200,000-dalton antigen present in extracts of RWP-1 and RWP-2 cells. This monoclonal antibody may be useful in the classification of pancreatic tumor cells.

It is the aim of this study to characterize and quantify the cells within isolated rat islets that express MHC class II antigens. A set of five monoclonal antibodies and two polyclonal antisera of defined specificity were used in combination with a newly devised procedure for three-dimensional immunofluorescence evaluation of intact islets. It is shown that in addition to passenger cells, such as lymphocytes, macrophages, and dendriticlike cells, vascular endothelial and endocrine cells are also capable of expressing class II antigens. This expression is strongly influenced by in vitro culture, pregnancy, streptozotocin-induced diabetes, transplantation trauma, and alloantigenic stimuli. The possible role of the above cells in antigen presentation related to islet transplantation is discussed.

We have reported a marked decrease of lymphocyte sensitivity to prostaglandins (PGs) in patients with untreated cervical carcinoma and other solid neoplasias. Particularly, Ea and ME rosette formation was inhibited only slightly by PGs in all cancer patients studied, as compared with the high inhibition values observed in normal individuals (Clin Immunol Immunopathol, 1981). In the present work, the PG-induced inhibition of Ea and ME rosette formation has been studied in patients with stage 0 cervical carcinoma up to 180 days after hysterectomy. It has been found that in these patients the PG sensitivity increased gradually up to or higher than the levels seen in normal control subjects. However, the PG sensitivity of Ea is restored earlier after surgery, while ME show a delayed recovery. Since the PG sensitivities correlate well with the clinical status of patients, it is suggested that such sensitivity may represent a useful test for following patients with cervical carcinoma.

The recognition that the multiple functions of T-lymphocytes are accomplished by separate subpopulations of cells that can be delineated both functionally and by antisera directed against specific cell surface markers is one major advance in immunology. Our study was directed at enumerating lymphocyte subsets in cytomegalovirus (CMV) perinatal infections with monoclonal antibodies and cell flow cytometry. Eight patients ranging in age from 1 week to 2 1/2 months were studied. Except for one term neonate, all of the infants were less than 36 weeks of gestational age, with a mean of 31.5 weeks (range 27-36 weeks) at birth. The mean birth weight of the premature infants was 1,211 gm (range 870-2420 gm). Five patients were 3 weeks old when studied and were classified as congenital CMV by viral isolation. Three were older than 2 months and were classified as acquired CMV. One of the acquired CMV infants was followed from birth to 6 months of age and had normal lymphocyte numbers at 2 weeks with alterations appearing at 8 weeks. The results of these studies indicated that in congenital CMV there is a normal to high helper/suppressor ratio. On the contrary, in acquired CMV this ratio is profoundly depressed because of significant increase in suppressor T-lymphocytes with borderline normal helper T-lymphocytes. In the latter group, the ratio increased with time after the acute infection episode. Our results agree with reported data indicating that in adults with CMV there is a depression of the helper/suppressor ratio during the acute stage with a return toward normal values during convalescence.

Membrane surface antigens on peripheral blood lymphocytes were analyzed by a laser flow cytometry system using monoclonal antibodies. Four normal subjects among 1,998 normal and diseased groups were found whose lymphocytes were not detected by OKT4 monoclonal antibody, while Leu3a and KOLT-1, which were also specific for helper/inducer T-lymphocytes as OKT4, were present in these subjects. The cases with the defect of the OKT4 antigen suggest that there is some modification of the antigen, lack of epitopes, or genetic polymorphism in the appearance of the OKT4+ cell surface antigen on normal human lymphocyte.

Preliminary data on a novel method for the diagnosis of bovine tuberculosis (TB) has been presented. This method is based on increased amino acid transport in specifically stimulated lymphocyte cultures. Lymphocytes isolated from TB-infected or -sensitized cattle and cultured with the TB antigen PPD-B had noticeably enhanced amino acid transport after 1-3 days. Transport continued to increase for up to 5 days of culture with the antigen. PPD-B did not stimulate transport in lymphocytes from nonsensitized animals. The new method correlated well with the tuberculin skin test and antigen-induced lymphocyte proliferation measured by flow cytometry.

The aim of the present study was to investigate the phagocytic response of normal human polymorphonuclear leukocytes (PMN) and monocytes (MN) to eight serotypes of C trachomatis (B,C,D,E,F,I,J, and L2) using a chemiluminescence (CL) assay, with luminal and lucigenin as amplifiers. The magnitude of the phagocytic cell CL response was proportional to the phagocyte-to-chlamydiae ratio, with a poor CL response detected at a ratio of 1:125 and progressively larger CL responses up to ratios of 1:50,000. The durations of the CL responses to all chlamydiae serotypes tested were considerably longer than that for zymosan. The PMN demonstrated a relatively greater CL response to all chlamydiae serotypes tested when compared with MN. The PMN and MN CL responses to "genital serotypes" (D,E,F,I, and J) (as well as lymphogranuloma venereum serotype L2) were greater than that for "ocular" serotypes (B and C). Inactivation of serum complement and specific chlamydial antibody absorption reduced the CL responses of both PMN and MN. This is the first study to characterize the CL responses of normal human PMN and MN cells to C trachomatis, and it indicates the important role of oxygen dependent antimicrobial systems in the phagocytosis of this common human pathogen.

Natural cell-mediated cytotoxicity (CYT) was determined using whole blood samples from 32 patients with the acquired immunodeficiency syndrome (AIDS) and 12 with AIDS-related complexes (ARC) and correlated with the number of putative natural killer cells (NK) bearing surface marker Leu 11a. Mean percent cytotoxicity (%CYT) was significantly lower in AIDS and ARC patients compared to that of normal controls. Mean %NK cells in AIDS patients was significantly higher than in normals, while %NK cells in ARC patients did not differ significantly from that of normals. In contrast, the absolute number of NK (NNK) cells in AIDS patients did not differ significantly from that of normals, while the NNK cells in ARC patients was significantly lower than those in normals. When the data were expressed in kinetic lytic units per NK cell (KLU/NK = maximum number of K562 cells killed per NK cell in 4 hours), significantly lower KLU/NK were observed in both AIDS and ARC patients compared to that in normals. Our data show that NK cells in AIDS and ARC patients are in a less active state compared to those in normals.